Suppression of ACE2 SUMOylation protects against SARS-CoV-2 infection through TOLLIP-mediated selective autophagy

In addition to investigating the virology of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), discovering the host–virus dependencies are essential to identify and design effective antiviral therapy strategy. Here, we report that the SARS-CoV-2 entry receptor, ACE2, conjugates with small ubiquitin-like modifier 3 (SUMO3) and provide evidence indicating that prevention of ACE2 SUMOylation can block SARS-CoV-2 infection. E3 SUMO ligase PIAS4 prompts the SUMOylation and stabilization of ACE2, whereas deSUMOylation enzyme SENP3 reverses this process. Conjugation of SUMO3 with ACE2 at lysine (K) 187 hampers the K48-linked ubiquitination of ACE2, thus suppressing its subsequent cargo receptor TOLLIP-dependent autophagic degradation. TOLLIP deficiency results in the stabilization of ACE2 and elevated SARS-CoV-2 infection. In conclusion, our findings suggest selective autophagic degradation of ACE2 orchestrated by SUMOylation and ubiquitination as a potential way to combat SARS-CoV-2 infection.


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We require information from authors about some types of materials, experimental systems and methods used in many studies. Here, indicate whether each material, system or method listed is relevant to your study. If you are not sure if a list item applies to your research, read the appropriate section before selecting a response.  Antibodies used proteomecentral.proteomexchange.org/cgi/GetDataset?ID=PXD034168]. The RNA-seq data have been deposited in public Gene Expression Omnibus (GEO) database under the accession number GSE171130 [https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE171130]. The whole-genome sequence (WGS) of SARS-CoV-2 isolate used in this study can be found with GenBank accession number MT123290. Source data are provided with this paper.
Sample size for each experiment is indicated in the legend. No statistical methods were used to predetermine sample sizes. Sample size was chosen based on previous experiments and comparable. The reference of cellular experiments sample size is: Jin, S.H. et al. Tetherin suppresses type I interferon signaling by targeting MAVS for NDP52-mediated selective autophagic degradation in human cells. Mol. Cell 68, 308-322 (2017).
No exclusion of data was made.
All experimental findings were reproduced in multiple independent experiments. For each figure, the number of independent experiments or biological replicates is indicated in the figure legends. Western blot and microscopy pictures are from a representative experiment and the number of independent repeats is clearly indicated in the figure legends.
For cell culture experiments, cells were split, plated in culture vessels, and then treated with DMSO or drugs. Because control and treatment groups were derived from the same cell line, no randomization could be performed. In animal study, mice were randomly allocated for each group.
Investigators were blinded to group allocation during data collection. Investigators were blinded for analysis of immunofluorescence staining. In experiments without subjective estimation like western blotting and qPCR, investigators were unblinded since no bias would be introduced by the investigators.
HEK293T, Calu-3, HeLa, A549, HepG2 and Vero E6 cells were authenticated for STR DNA profiling by National Collection of Authenticated Cell Cultures (Shanghai, China). hUVECs were authenticated for STR DNA profiling by China Center for Type Culture Collection (Wuhan, China), no further authentication performed in the laboratory.
These cell lines have been tested for mycoplasma contamination by MycoAler Mycoplasma Detection Kit (R&D Systems, cat. CUL001B) and the results of detection showed that cultured cells were not contaimated by mycoplasma.
There is no any commonly misidentified cell lines in this study.